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Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometric (LC/ESI-MS/MS) Study for the Identification and Characterization of In Vivo Metabolites of Cisplatin in Rat Kidney Cancer Tissues: Online Hydrogen/Deuterium (H/D) Exchange Study.

Bandu R, Ahn HS, Lee JW, Kim YW, Choi SH, Kim HJ, Kim KP - PLoS ONE (2015)

Bottom Line: A total of thirty one unknown in vivo metabolites have been identified and the structures of metabolites were elucidated using LC-MS/MS experiments combined with accurate mass measurements.Online HDX experiments have been used to further support the structural characterization of metabolites.The results showed that CP undergoes a series of ligand exchange biotransformation reactions with water and other nucleophiles like thio groups of methionine, cysteine, acetylcysteine, glutathione and thioether.

View Article: PubMed Central - PubMed

Affiliation: Department of Applied Chemistry, College of Applied Sciences, Kyung Hee University, Yong-in City, Republic of Korea.

ABSTRACT
In vivo rat kidney tissue metabolites of an anticancer drug, cisplatin (cis-diamminedichloroplatinum [II]) (CP) which is used for the treatment of testicular, ovarian, bladder, cervical, esophageal, small cell lung, head and neck cancers, have been identified and characterized by using liquid chromatography positive ion electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) in combination with on line hydrogen/deuterium exchange (HDX) experiments. To identify in vivo metabolites, kidney tissues were collected after intravenous administration of CP to adult male Sprague-Dawley rats (n = 3 per group). The tissue samples were homogenized and extracted using newly optimized metabolite extraction procedure which involves liquid extraction with phosphate buffer containing ethyl acetate and protein precipitation with mixed solvents of methanol-water-chloroform followed by solid-phase clean-up procedure on Oasis HLB 3cc cartridges and then subjected to LC/ESI-HRMS analysis. A total of thirty one unknown in vivo metabolites have been identified and the structures of metabolites were elucidated using LC-MS/MS experiments combined with accurate mass measurements. Online HDX experiments have been used to further support the structural characterization of metabolites. The results showed that CP undergoes a series of ligand exchange biotransformation reactions with water and other nucleophiles like thio groups of methionine, cysteine, acetylcysteine, glutathione and thioether. This is the first research approach focused on the structure elucidation of biotransformation products of CP in rats, and the identification of metabolites provides essential information for further pharmacological and clinical studies of CP, and may also be useful to develop various effective new anticancer agents.

No MeSH data available.


Related in: MedlinePlus

Proposed fragmentation mechanism for metabolite M15.
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pone.0134027.g010: Proposed fragmentation mechanism for metabolite M15.

Mentions: M15 ([M+H]+; m/z 528.4816): The second most abundant metabolite M15 at m/z 528.4816 ([M+H]+) with an elemental composition of C10H29N4O4PtS2 (3.96 ppm) was eluted at 14.6 min. The HRMS data suggests the lack of one chlorine atom and presence of an additional methionine moiety in M15 as compared to M5. This can be seen from the LC-MS/MS spectrum of protonated M15 which shows abundant product ions at m/z 378.4711 and m/z 228.4812 corresponding to a probable loss of one and two methionine moieties, respectively, and m/z 150.0587 (protonated methionine) (Fig 10). It can be noted that the peaks at m/z 264.4732 (Pt+(NH3)2Cl) and m/z 246.5177 (Pt+(NH3)2OH) would have been present and the sequential losses of two neutral species of methionine would have been absent in case of one methionine moiety in protonated M15 as discussed in M5 and M6. Further support comes from the appearance of moderately abundant product ions at m/z 482.4733 and m/z 436.4822 which were formed by the neutral loss of one and two formic acids, respectively, from protonated M15. Similarly to M5 and M6, the appearance of m/z 150.0587 ion (protonated methionine) in the MS/MS of protonated M15 and m/z 154.0816 ion (deuterated methionine) in the MS/MS of deuterated M15, reflects the presence of methionine moiety in the structure of M15. Based on these data, the structure of M15 can be assigned as a di-methionine metabolite of CP.


Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometric (LC/ESI-MS/MS) Study for the Identification and Characterization of In Vivo Metabolites of Cisplatin in Rat Kidney Cancer Tissues: Online Hydrogen/Deuterium (H/D) Exchange Study.

Bandu R, Ahn HS, Lee JW, Kim YW, Choi SH, Kim HJ, Kim KP - PLoS ONE (2015)

Proposed fragmentation mechanism for metabolite M15.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4526507&req=5

pone.0134027.g010: Proposed fragmentation mechanism for metabolite M15.
Mentions: M15 ([M+H]+; m/z 528.4816): The second most abundant metabolite M15 at m/z 528.4816 ([M+H]+) with an elemental composition of C10H29N4O4PtS2 (3.96 ppm) was eluted at 14.6 min. The HRMS data suggests the lack of one chlorine atom and presence of an additional methionine moiety in M15 as compared to M5. This can be seen from the LC-MS/MS spectrum of protonated M15 which shows abundant product ions at m/z 378.4711 and m/z 228.4812 corresponding to a probable loss of one and two methionine moieties, respectively, and m/z 150.0587 (protonated methionine) (Fig 10). It can be noted that the peaks at m/z 264.4732 (Pt+(NH3)2Cl) and m/z 246.5177 (Pt+(NH3)2OH) would have been present and the sequential losses of two neutral species of methionine would have been absent in case of one methionine moiety in protonated M15 as discussed in M5 and M6. Further support comes from the appearance of moderately abundant product ions at m/z 482.4733 and m/z 436.4822 which were formed by the neutral loss of one and two formic acids, respectively, from protonated M15. Similarly to M5 and M6, the appearance of m/z 150.0587 ion (protonated methionine) in the MS/MS of protonated M15 and m/z 154.0816 ion (deuterated methionine) in the MS/MS of deuterated M15, reflects the presence of methionine moiety in the structure of M15. Based on these data, the structure of M15 can be assigned as a di-methionine metabolite of CP.

Bottom Line: A total of thirty one unknown in vivo metabolites have been identified and the structures of metabolites were elucidated using LC-MS/MS experiments combined with accurate mass measurements.Online HDX experiments have been used to further support the structural characterization of metabolites.The results showed that CP undergoes a series of ligand exchange biotransformation reactions with water and other nucleophiles like thio groups of methionine, cysteine, acetylcysteine, glutathione and thioether.

View Article: PubMed Central - PubMed

Affiliation: Department of Applied Chemistry, College of Applied Sciences, Kyung Hee University, Yong-in City, Republic of Korea.

ABSTRACT
In vivo rat kidney tissue metabolites of an anticancer drug, cisplatin (cis-diamminedichloroplatinum [II]) (CP) which is used for the treatment of testicular, ovarian, bladder, cervical, esophageal, small cell lung, head and neck cancers, have been identified and characterized by using liquid chromatography positive ion electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) in combination with on line hydrogen/deuterium exchange (HDX) experiments. To identify in vivo metabolites, kidney tissues were collected after intravenous administration of CP to adult male Sprague-Dawley rats (n = 3 per group). The tissue samples were homogenized and extracted using newly optimized metabolite extraction procedure which involves liquid extraction with phosphate buffer containing ethyl acetate and protein precipitation with mixed solvents of methanol-water-chloroform followed by solid-phase clean-up procedure on Oasis HLB 3cc cartridges and then subjected to LC/ESI-HRMS analysis. A total of thirty one unknown in vivo metabolites have been identified and the structures of metabolites were elucidated using LC-MS/MS experiments combined with accurate mass measurements. Online HDX experiments have been used to further support the structural characterization of metabolites. The results showed that CP undergoes a series of ligand exchange biotransformation reactions with water and other nucleophiles like thio groups of methionine, cysteine, acetylcysteine, glutathione and thioether. This is the first research approach focused on the structure elucidation of biotransformation products of CP in rats, and the identification of metabolites provides essential information for further pharmacological and clinical studies of CP, and may also be useful to develop various effective new anticancer agents.

No MeSH data available.


Related in: MedlinePlus